ArticleJournal of virology2003
Determination of infectious retrovirus concentration from colony-forming assay with quantitative analysis.
Article in Journal of virology, 2003. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 27 papers.
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Who cites it
27 citing papers in PubMed, 68 citations in OpenAlex.
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- Role of Porphyromonas gingivalis outer membrane vesicles in oral mucosal transmission of HIV.Scientific reports · 2018Article
- Early HIV infection predictions: role of viral replication errors.SIAM journal on applied mathematics · 2018Article
- HIV dynamics linked to memory CD4+ T cell homeostasis.PloS one · 2017Article
- Optical manipulation of a single human virus for study of viral-cell interactions.Proceedings of SPIE--the International Society for Optical Engineering · 2016Article
- Preparation of MS2 Phage-Like Particles and Their Use As Potential Process Control Viruses for Detection and Quantification of Enteric RNA Viruses in Different Matrices.Frontiers in microbiology · 2016Article
- Recombination Enhances HIV-1 Envelope Diversity by Facilitating the Survival of Latent Genomic Fragments in the Plasma Virus Population.PLoS computational biology · 2015Article
- Antigenic properties of the human immunodeficiency virus envelope glycoprotein gp120 on virions bound to target cells.PLoS pathogens · 2015Article
- Molecular determinants of the ratio of inert to infectious virus particles.Progress in molecular biology and translational science · 2015Review
- Overexpression of the PLAP-1 gene inhibits the differentiation of BMSCs into osteoblast-like cells.Journal of molecular histology · 2014Article
- Basis for early and preferential selection of the E138K mutation in HIV-1 reverse transcriptase.Antimicrobial agents and chemotherapy · 2013Article
- Optimized Infectivity of the Cell-Free Single-Cycle Human Immunodeficiency Viruses Type 1 (HIV-1) and Its Restriction by Host Cells.PloS one · 2013Article
- Quantification system for the viral dynamics of a highly pathogenic simian/human immunodeficiency virus based on an in vitro experiment and a mathematical model.Retrovirology · 2012Article
- Conditional expression of the TVA receptor allows clonal analysis of descendents from Cre-expressing progenitor cells.Developmental biology · 2011Article
- Stochastic theory of early viral infection: continuous versus burst production of virions.PLoS computational biology · 2011Article
- Multilineage differentiation of dental follicle cells and the roles of Runx2 over-expression in enhancing osteoblast/cementoblast-related gene expression in dental follicle cells.Cell proliferation · 2010Article
- Expression and immunological analysis of capsid protein precursor of swine vesicular disease virus HK/70.Virologica Sinica · 2010Article
- Viral dynamics during primary simian immunodeficiency virus infection: effect of time-dependent virus infectivity.Journal of virology · 2010Article
- Rapid dissociation of HIV-1 from cultured cells severely limits infectivity assays, causes the inactivation ascribed to entry inhibitors, and masks the inherently high level of infectivity of virions.Journal of virology · 2010Article
Corrections and comments
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Authors and funding
5 authors at 2 institutions in 2 countries.
Funding
Abstract
The colony formation assay is the most commonly used titration method for defining the concentration of replication-incompetent murine leukemia virus-derived retroviral vectors. However, titer varies with target cell type and number, transduction time, and concentration of polycation (e.g., Polybrene). Moreover, because most of the viruses cannot encounter target cells due to Brownian motion, their short half-lives, and the requirement for target cell division for activity, the actual infectious retrovirus concentration in the collected supernatant is higher than the viral titer. Here we correlate the physical viral particle concentration with the infectious virus concentration and colony formation titer with the help of a mathematical model. Ecotropic murine leukemia retrovirus supernatant, collected from the GP+E86/LNCX retroviral vector producer cell line, was concentrated by centrifugation and further purified by a sucrose density gradient. The physical concentration of purified viral vectors was determined by direct particle counting with an electron microscope. The concentrations of fresh and concentrated supernatant were determined by a quantitative reverse transcriptase activity assay. Titration of all supernatants by neomycin-resistant colony formation assay was also performed. There were 767 +/- 517 physical viral particles per infectious CFU in the crude viral supernatant. However, the infectious viral concentration determined by mathematical simulation was 143 viral particles per infectious unit, which is more consistent with the concentration determined by particle counting in purified viral solution. Our results suggest that the mathematical model can be used to extract a more accurate and reliable concentration of infectious retrovirus.
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